CN107073032B - Renal failure development inhibitor, renal failure preventive agent, and indoxyl sulfate production inhibitor - Google Patents

Abstract

The present invention provides a novel inhibitor of renal failure progression, a renal failure prophylactic agent, and an indoxyl sulfate production inhibitor for humans, particularly renal failure patients. The inhibitor for renal failure development, the prophylactic agent for renal failure and the inhibitor for indoxyl sulfate production contain paramylon derived from Euglena or processed product thereof as an active ingredient. For example, in the case of an inhibitor of renal failure progression, 1-5 g of paramylon is administered orally several times per 1 day to a patient who has chronic renal failure and is undergoing dialysis therapy. In particular, the administration is carried out separately in the form of a capsule or powder, with a predetermined time between the administration of the other drug and the administration of the other drug.

Description

Renal failure development inhibitor, renal failure preventive agent, and indoxyl sulfate production inhibitor

technical field

The present invention relates to a novel renal failure development inhibitor, renal failure preventive agent and indoxyl sulfate production inhibitor.

Background technique

Currently, there are more than 300,000 patients with end-stage renal failure in Japan, and the number is still increasing. There is no fundamental treatment for end-stage renal failure. As the main renal replacement therapy, there are hemodialysis, peritoneal dialysis and kidney transplantation. However, any alternative therapy brings a great burden to the patient. Therefore, it is important to suppress the progression of renal failure at the initial stage of symptoms, and it is expected that there are effective renal failure progression inhibitors and renal failure therapeutic agents.

It is generally known that in renal failure patients, the following values are increased: the blood concentration of indoxyl sulfate, a uremic substance that is a factor for the development of renal failure or a vascular disorder factor, and the blood level of creatinine, which is an indicator of renal dysfunction concentration, blood concentration of homocysteine, which is a risk factor for cardiovascular disease caused by atherosclerosis, and neutral fat.

Among them, it has been reported that indoxyl sulfate is a uremic substance proven to be a development factor for renal failure (see Non-Patent Document 1), and that the concentration of indoxyl sulfate in the blood of patients with chronic renal failure is higher than that of healthy subjects. Abnormally high concentrations of dooxyl.

As described above, since indoxyl sulfate is a substance that aggravates and worsens renal failure, it is considered that by reducing the blood indoxyl sulfate concentration of renal failure patients, it is possible to significantly reduce renal dysfunction and inhibit the progression of renal failure. On the other hand, indoxyl sulfate is a difficult metabolite that cannot be removed even by the above-mentioned renal replacement therapy.

For this reason, it is known that spherical adsorbent carbon, which is a conventional inhibitor of the progression of renal failure, adsorbs indole, a precursor of indoxyl sulfate, in the intestine, and excretes it into feces, thereby reducing the concentration of indoxyl sulfate in the blood. . As a result, it has been reported that in addition to suppressing the progression of renal failure, the onset of cardiovascular disease related to renal failure is also suppressed, and the mortality associated with cardiovascular disease is also reduced.

However, in the case of spherical adsorbent carbon, oral administration is burdened for reasons such as having to take 30 capsules per day, and symptoms such as abdominal distention and constipation as side effects are severe. Painful, so it is confirmed that many patients want to stop taking spherical adsorption charcoal.

Therefore, there is a need for novel agents that reduce the pain and burden of patients with a smaller dosage without causing abdominal distention, constipation, and the like.

As a possibility of a novel drug, for example, a chronic renal failure preventive and therapeutic agent containing chitosan, which is known as an adsorbent, has been proposed (see Patent Document 1).

Specifically, it has been reported that, based on the results of animal experiments using renal failure model rats, oral administration of chitosan as an active ingredient has shown that a uremic substance, that is, sulfuric acid, which is a causative substance that causes an oxidative stress environment, The blood concentration of indoxyl decreases.

However, the results of clinical trials on humans, especially patients with chronic renal failure, have not been reported. In order to verify the efficacy and safety of the patients, or to reduce the pain and burden of these patients, it is necessary to administer drugs. The usage or dosage of time, administration procedure, administration amount, etc. is further studied.

On the other hand, Euglena (generic name: Euglena, Japanese name: ミドリムシ) is attracting attention as a biological resource expected to be used as food, feed, fuel, and the like.

Euglena contains 59 nutrients, including vitamins, minerals, amino acids, unsaturated fatty acids, etc., which are equivalent to most of the nutrients required for human survival. Possibility to utilize food supply sources in poor areas that consume essential nutrients.

Euglena is located at the bottom of the food chain, and it is difficult to cultivate in large quantities because it is preyed on by predators and the cultivation conditions such as light, temperature conditions, and stirring speed are difficult compared with other microorganisms. However, in recent years, the present inventors have established through intensive research. Mass culture technology has opened the way of massive supply of Euglena and paramylon extracted from Euglena.

Euglena is a unique organism that has animal properties that carry out flagellar movement, and simultaneously performs photosynthesis as a plant having chloroplasts, and Euglena itself and substances derived from Euglena are expected to have various functions.

Therefore, elucidation of the functions and functional expression mechanisms of substances derived from Euglena, such as Euglena and paramylon, which are already available in large quantities, and development of methods for utilizing these substances are desired.

prior art literature

Patent Literature

Patent Document 1: Japanese Patent Laid-Open No. 2014-24817

Non-patent literature

Non-Patent Document 1: Niwa Ritsu, Modern Medicine Vol. 47 No. 1: 55-61 (1999)

Non-Patent Document 2: Tanaka, R. et al.: Jpn. J. Pediatr., 33, 2483 (1980)

SUMMARY OF THE INVENTION

The problem to be solved by the invention

The present invention has been accomplished in view of the above-mentioned problems, and an object of the present invention is to provide a novel renal failure progression inhibitor, renal failure preventive agent, and indoxyl sulfate production inhibitor for living organisms.

Another object of the present invention is to provide a renal failure progression inhibitor, a renal failure preventive agent, and indole sulfate that reduce the blood concentration of indoxyl sulfate, which is difficult to remove even by dialysis therapy in humans, especially in patients with renal failure. Phenol production inhibitor.

Another object of the present invention is to provide a renal failure development inhibitor, a renal failure preventive agent, and an indoxyl sulfate production inhibitor, which are novel methods of utilizing a Euglena-derived substance.

means of solving problems

As a result of intensive studies, the present inventors found that the blood concentration of indoxyl sulfate, which is a uremic substance, decreases when paramylon derived from Euglena algae is administered to a living body.

In detail, indole, the precursor of indoxyl sulfate, is a spoilage product produced by the decomposition of tryptophan contained in a large amount of protein in food by harmful bacteria in the intestine. When paramylon or a paramylon processed product is administered to a failure patient, the paramylon adsorbs indole, and the concentration of indoxyl sulfate in the blood decreases, thereby completing the present invention.

It is also known that in general, in the intestinal tract of patients undergoing artificial dialysis of blood due to renal failure, harmful intestinal bacteria such as coliforms increase, while anaerobic bacteria beneficial to the human body such as bifid lactic acid bacteria and lactic acid bacteria decrease. Furthermore, it was confirmed that intestinal flora was improved by ingesting bifid lactic acid bacteria, lactic acid bacteria, and the like (Non-Patent Document 2).

Therefore, as a result of intensive research by the present inventors, it has been found that paramylon derived from Euglena can contribute to the improvement of the human intestinal environment. Specifically, when paramylon derived from Euglena algae is administered to a living body, harmful bacteria in the intestines are significantly reduced and bifidobacteria and lactic acid bacteria in the intestines are increased, thereby completing the present invention.

In addition, as a result of intensive research by the present inventors, it has been found and clarified that when paramylon derived from Euglena algae is administered to a living body, there is a synergistic effect not seen in conventional spherical adsorbent carbons in the intestines of orally ingested food. By shortening the time, the amount of indole produced by the decomposition of tryptophan in the intestine is suppressed, thereby completing the present invention.

Therefore, the said subject is solved by the following means, that is, the renal failure progression inhibitor of this invention contains paramylon or its processed product as an active ingredient.

According to the above configuration, when paramylon or its processed product is administered to a patient with renal failure, paramylon reduces the concentration of indoxyl sulfate in the blood, so the present invention can be used as a development inhibitor of renal failure disease.

At this time, it is appropriate to administer the drug to patients with renal failure who are receiving dialysis therapy.

According to the above configuration, indoxyl sulfate is generally a uremic substance that is difficult to remove even by dialysis therapy. By administering paramylon to a patient undergoing dialysis therapy, the blood concentration of indoxyl sulfate in the patient can be reduced, Further exacerbation and deterioration of, for example, end-stage renal failure can be prevented.

In this case, it is appropriate to administer the drug to a patient who has chronic renal failure and is 50 to 70 years old.

According to the above structure, the average age of patients with end-stage renal failure who are generally introduced to dialysis therapy is 68.4 years old (current status of chronic dialysis therapy in Japan as of December 31, 2012, Japan Dialysis Medicine Association). Paramylon is administered to patients aged 50 to 70 years in the pre-stage, so that the development of renal failure in the patient can be prevented in the pre-stage of the introduction of dialysis therapy.

In this case, it is appropriate to continuously administer 1 to 5 g of the paramylon or its processed product to a patient suffering from chronic renal failure three times a day.

In addition, oral administration in the form of capsules or powders is suitable for patients suffering from chronic renal failure.

In addition, it is suitable to give a single drug to a patient suffering from chronic renal failure with a predetermined period of time before and after the administration of other drugs.

According to the above configuration, the administration time, administration procedure, administration amount, etc. of paramylon with high effectiveness are limited to the patients with renal failure, and on this basis, paramylon or its processing can be provided. An inhibitor of renal failure development as an active ingredient.

In addition, a renal failure preventive agent, a uremia therapeutic agent, an indoxyl sulfate production inhibitor, or a cardiovascular system disease preventive agent containing paramylon or a processed product thereof as an active ingredient can also be realized.

In addition, a specific health food for suppressing the progression of renal failure containing paramylon or a processed product thereof as an active ingredient can also be realized.

In addition, a method for suppressing the development of renal failure by ingesting a composition containing paramylon or a processed product thereof as an active ingredient can also be realized (excluding medical action on humans).

Furthermore, a method for inhibiting the development of renal failure including the step of administering or ingesting an effective amount of paramylon or a processed product thereof to a living body (human), particularly a patient, can also be realized.

In addition, the use of paramylon or its processed product for the manufacture of a drug for suppressing the progression of renal failure can also be realized.

In addition, the use of the renal failure development inhibiting substance characterized in that the renal failure development inhibiting substance is paramylon or a processed product thereof for the manufacture of a drug for inhibiting the development of renal failure can also be realized.

Invention effect

According to the present invention, a novel renal failure progression inhibitor, renal failure preventive agent, and indoxyl sulfate production inhibitor can be provided.

In addition, it is possible to provide a renal failure progression inhibitor, a renal failure preventive agent, and an indoxyl sulfate production inhibitor that reduce the blood concentration of indoxyl sulfate, which is difficult to remove even by dialysis therapy in humans, especially in patients with renal failure. .

In addition, it is possible to provide a kidney failure development inhibitor, a kidney failure preventive agent, and an indoxyl sulfate production inhibitor, which are novel utilization methods of Euglena-derived substances.

Description of drawings

FIG. 1 is a graph showing the amount of change in blood indoxyl sulfate concentration when the renal failure progression inhibitor of this example was administered to renal failure patients for 9 weeks.

2 is a graph showing the amount of change in blood indoxyl sulfate concentration when a renal failure progression inhibitor is administered to renal failure patients aged 50 years or older and under 70 years old for 9 weeks.

3 is a graph showing the amount of change in blood indoxyl sulfate concentration when a renal failure progression inhibitor is administered to a male renal failure patient for 9 weeks.

FIG. 4 is a graph showing a time-dependent change in the residual concentration of indole in the solution when paramylon and an indole solution of the present example were mixed.

5 is a graph comparing the occupancy rates of intestinal flora in the cecum when paramylon according to an example of the present invention is administered to rats for 4 weeks.

FIG. 6 is a graph comparing the ratios of beneficial bacteria/harmful bacteria in the cecum in FIG. 5 .

FIG. 7 is a graph comparing the intestinal transit time in the second week and the fourth week from the start of feeding when the paramylon of the present Example was administered to the rat for four weeks.

8 is a graph comparing the weight of feces in the second and fourth weeks from the start of feeding when the rats were administered paramylon according to this example for four weeks.

FIG. 9 is a graph comparing the water content of feces in the second week and the fourth week from the start of rearing in FIG. 8 .

Detailed ways

Hereinafter, embodiments of the present invention will be described with reference to FIGS. 1 to 9 .

The present embodiment relates to a renal failure progression inhibitor, which contains paramylon derived from Euglena or a processed product thereof as a main component, and is administered to a renal failure patient, thereby increasing the concentration of indoxyl sulfate in the blood of the living body. Reduce, inhibit the development and deterioration of renal failure diseases.

<Outline of kidney failure diseases>

Renal failure refers to a state in which renal function is reduced to about 30% or less of normal, and includes acute renal failure in which renal function is rapidly decreased and chronic renal failure in which renal function is gradually decreased over many years.

First of all, acute renal failure refers to: the result of a sharp decline in renal function, resulting in high blood creatinine concentration (for example, an increase of 0.5 mg/dL or more per day) and high blood urea nitrogen concentration (for example, a daily increase of 10 mg/dL). dL or more), moisture in body fluids, abnormal electrolyte concentrations, etc., and are in a state where the constancy of body fluids cannot be maintained.

Acute renal failure is classified into prerenal, renal and postrenal depending on the location of the cause.

Prerenal acute renal failure is a pathological condition in which renal blood flow decreases and glomerular filtration rate decreases due to deterioration of blood circulation dynamics. The main causes include heart failure, myocardial infarction, epicarditis, and vasculitis. , arteriosclerosis, bilateral renal artery stenosis, activation of the renin-angiotensin system, sepsis, allergies, cirrhosis, anesthetics, bleeding, dehydration, vomiting, diarrhea, edema, ascites, burns, nephrotic syndrome symptoms, adrenal insufficiency, etc.

Acute renal failure is a pathological condition caused by disturbance of the glomeruli, tubules, and interstitium of the kidney, and acute glomerulonephritis, collagen disease, hemolytic uremic syndrome, acute tubular Necrosis, hypercalcemia, drug allergy, pyelonephritis, NSAIDs, acute tubular necrosis in the narrow sense from prerenal passage, antibiotics, contrast agents, heavy metals, multiple myeloma, hyperuricemia, rhabdomyolysis, DIC Wait.

Postrenal acute renal failure is a morbid state caused by obstruction of the urinary tract, and ureteral obstruction (calculus, tumor, retroperitoneal fibrosis), prostatic hypertrophy, tumor, calculus, etc. are mentioned as the main cause.

Acute renal failure generally requires dialysis therapy. If dialysis therapy is effective, it will go through the onset period, oliguria period, diuretic period, and recovery period in turn to achieve cure. On the other hand, in severe cases, there may be cases where the kidney function cannot be restored and the dialysis treatment is continued.

Although the mortality rate of acute renal failure has been reduced due to the advancement of dialysis therapy, the mortality rate is still about 50%, and it is also a dangerous disease in renal disease.

Next, chronic renal failure refers to a pathological condition in which kidney function is irreversibly gradually decreased due to various chronically advanced kidney diseases, and the constant of body fluids cannot be maintained, and various pathological conditions such as hypertension, anemia and abnormal bone metabolism are exhibited. In particular, many symptoms of end-stage renal failure are called uremia, and dialysis therapy is required to purify the blood.

The development of chronic renal failure is generally divided into stages 1 to 5 according to the disease classification of chronic kidney disease (CKD: Chronic Kidney Disease) (CKD diagnosis and treatment guidelines 2012, Nippon Kidney Association 2012 (CKD diagnosis and treatment ガイド 2012, Nippon kidney disease 2012) Society Chi 2012)).

Chronic kidney disease (CKD) is diagnosed by proteinuria and glomerular filtration rate ((GFR) ml/min/1.73m ² ) in daily clinical practice. Age and sex were evaluated using estimated GFR (eGFR) calculated using the Japanese GFR estimation formula.

It should be noted that the glomerular filtration rate (GFR) is an index for measuring the quality of renal function (the ability to excrete waste in the form of urine), and the lower the value, the lower the renal function.

The first stage of CKD is a state with a normal GFR value (GFR≥90) despite renal impairment, and the second stage is a state with renal impairment and a mild GFR value (GFR=60 to 89). During this stage, residual renal function is functioning and is largely asymptomatic.

The third stage of CKD is a state with renal impairment and a moderate GFR value (GFR=30 to 59), the residual renal function is not complete, an increase in urine volume and an increase in blood urea nitrogen are confirmed, and there are also Mild anemia, unable to maintain the constancy of body fluids.

The fourth stage of CKD is a state in which there is renal impairment and a high GFR value (GFR=15 to 29), and the symptoms of the third stage are aggravated. In addition, the 1st - 4th stage of CKD is also called preservation period (original text: preservation period) renal failure in the state before the start of dialysis therapy.

Then, the fifth stage of CKD is a state of end-stage renal failure requiring dialysis therapy (GFR<15), and symptoms of uremia appear along with aggravation of abnormal body fluids.

Diabetic nephropathy, chronic nephritis (chronic glomerulonephritis), nephrosclerosis, etc. are mentioned as main diseases which cause chronic renal failure.

In the case of end-stage renal failure, if the kidney function is not replaced by dialysis therapy or a kidney transplant is performed, the person is at the risk of dying. The standard for starting dialysis therapy is generally a blood creatinine value of 8 mg/dl or more or a blood urea nitrogen value of 100 mg/dl or more.

It should be noted that dialysis therapy refers to a therapy method for artificially performing dialysis to purify blood when wastes cannot be removed due to renal failure or uremia, and mainly includes two types of hemodialysis and peritoneal dialysis.

In addition, hemofiltration for excellent removal of medium molecular weight substances by hemodialysis, hemofiltration dialysis for excellent removal of substances ranging from small molecular substances to low protein substances by combining hemodialysis and hemofiltration, and hemofiltration dialysis for the reduction of cardiac function For patients who cannot tolerate hemodialysis, continuous hemofiltration dialysis, which is slowly and time-consuming, and hemoadsorption in which specific substances are removed using adsorbents, and plasma exchange in which plasma is separated and removed from blood and replenished with new plasma, etc. Law.

<Outline of indoxyl sulfate>

Next, indoxyl sulfate, which is a uremic substance that exhibits high blood levels in renal failure patients and is an aggravating factor of renal failure disease, will be described.

Indoxyl sulfate is a metabolite of protein in food. Specifically, tryptophan contained in protein is decomposed by intestinal harmful bacteria such as Escherichia coli, thereby producing indole as a precursor. Then, after indole is absorbed by the digestive tract, it is combined with sulfuric acid in the liver to produce indoxyl sulfate.

Indoxyl sulfate is released into the blood and exists mostly in the form bound to albumin, which is not metabolized and is mainly excreted from the kidneys into the urine. A state of accumulation in high concentrations in blood.

Indoxyl sulfate is involved in renal fibrosis, glomerulosclerosis, and the like, and is known to induce the induction of reactive oxygen species, decrease free radical scavengers, cause cardiovascular disease, and develop and exacerbate renal failure.

In addition, it is known that the blood indoxyl sulfate concentration in the living body is related to the blood creatinine concentration and the blood urea nitrogen concentration, which are indicators of renal function. Impairment of renal function associated with failure was significantly reduced.

Therefore, it is considered to be involved in suppressing the progression of renal failure disease by reducing the blood indoxyl sulfate concentration in the body of renal failure patients.

＜Renal failure development inhibitor＞

The paramylon which is the main component of the renal failure development inhibitor or its processed product includes paramylon extracted from paramyel cells, paramylon powder, various processed products of paramylon, and the like.

As the Euglena cells, it is desirable to use Euglena gracilis (E. gracilis), especially E. gracilis Z strain. In addition, species such as Euglena gracilis Krebs, Euglena gracilis Barubachirasu, E. gracilis Z strain mutant strain SM-ZK strain (chloroplast-deficient strain), mutant var. bacillaris, and chloroplast mutant strains of these species may be used. β-1,3-glucanases derived from genetic variants, Euglena intermedia, Euglena piride and other Euglena species such as Astaia longa.

Euglena is widely distributed in fresh water such as ponds and swamps, and can be isolated and used, and any Euglena that has been isolated can also be used.

Euglena of the present invention includes all of its variant strains. In addition, products obtained by genetic methods such as recombination, transduction, transformation and the like are also included in these mutant strains.

In the culture of Euglena cells, as a culture medium, for example, a culture medium to which nutrients such as nitrogen sources, phosphorus sources, minerals, etc. are added, for example, an improved Cramer-Myers medium ((NH ₄ ) ₂ HPO ₄ 1.0 g can be used /L, KH ₂ PO ₄ 1.0 g/L, MgSO ₄ 7H ₂ O 0.2 g/L, CaCl ₂ 2H ₂ O 0.02 g/L, Fe ₂ (SO ₂ ) ₃ 7H ₂ O 3 mg/L, MnCl _{2.4H2O 1.8mg} /L, _{CoSO4.7H2O 1.5mg} /L, _{ZnSO4.7H2O 0.4mg} / _L , _{Na2MoO4.2H2O 0.2mg} / _L , _CuSO4.5H2 O 0.02 g/L, thiamine hydrochloride (vitamin B ₁ ) 0.1 mg/L, cyanocobalamin (vitamin B ₁₂ ), (pH 3.5)). In addition, (NH ₄ ) ₂ HPO ₄ may be changed to (NH ₄ ) ₂ SO ₄ or NH ₃ aq. In addition, a known Hutner medium and Koren-Hutner medium prepared based on the description of "Yu グ レ ナ Physiology and Biochemistry" (edited by Masaburo Kitaoka, Co., Ltd.) can also be used.

The pH of the culture solution is preferably 2 or more, and the upper limit thereof is preferably 6 or less, more preferably 4.5 or less. By setting pH on the acidic side, photosynthetic microorganisms can grow and develop more dominantly than other microorganisms, and thus contamination can be suppressed.

In addition, the culture of Euglena cells can be carried out, for example, by the intermittent feeding method, or by flask culture, culture using a fermenter, batch culture method, or semi-batch culture method (fed-batch culture method). , continuous culture method (perfusion culture method) and any other liquid culture method.

Separation of Euglena cells is performed, for example, by centrifugation of the culture solution or simple sedimentation.

Paramylon is a macromolecular body (β-1,3-glucan) in which about 700 glucoses are polymerized by β-1,3-bonds. polysaccharides. Paramylon particles are flat spheroid particles, and are formed by helically winding β-1,3-glucan chains.

Paramylon particles are isolated and purified from the cultured paramylon cells by any suitable method, and are usually provided in the form of powder.

For example, paramylon particles can be obtained by (1) culturing paramylon cells in any suitable medium; (2) isolating paramylon cells from the medium; (3) isolating paramylon from the isolated paramylon cells; (4) Purification of the isolated paramylon; and (5) cooling and subsequent freeze-drying as necessary.

The isolation of paramylon can be carried out using, for example, nonionic or anionic surfactants of mostly biodegradable species. The purification of paramylon can be performed substantially simultaneously with the separation.

In addition, isolation and purification of paramylon from Euglena are well known, and are described in, for example, E. Ziegler, "Die naturlichen und kunstlichen Aromen" Heidelberg, Germany, 1982, Chapter 4.3 "Gefriertrocken", DE 43 28 329 , or in JP 2003-529538.

As a processed product of paramylon, amorphous paramylon is mentioned, for example.

Amorphous paramylon refers to a substance obtained by amorphizing crystalline paramylon derived from Euglena.

The relative crystallinity of amorphous paramylon with respect to crystalline paramylon produced from Euglena by a known method is 1 to 20%.

Here, the relative crystallinity is obtained by the method described in Japanese Patent Application No. 2010-52042.

That is, amorphous paramylon and paramylon were pulverized for 5 minutes with a pulverizer (Ball Mill MM400 manufactured by Retsch Corporation) at a vibration frequency of 20 times/sec, respectively, and then subjected to an X-ray diffractometer (H'PertPRO manufactured by Specris Corporation) in the Under the condition of tube voltage 45KV and tube current 40mA, 2θ was scanned in the range of 5° to 30°, and the diffraction peaks Pc and Pa of paramylon and amorphous paramylon near 2θ=20° were obtained.

Using the values of Pc and Pa, pass

Relative crystallinity of amorphous paramylon=Pa/Pc×100(%)

The relative crystallinity of amorphous paramylon was calculated.

Amorphous paramylon can be prepared by subjecting crystalline paramylon powder to alkali treatment, neutralizing it with an acid, washing, removing moisture, and drying it according to the method described in Japanese Patent Application No. 2010-52042. preparation.

Processed products of paramylon include water-soluble paramylon, sulfated paramylon, etc., and paramylon derivatives obtained by chemically or physically treating paramylon by various known methods.

＜＜Inhibition of the development of renal failure＞

The renal failure progression inhibitor can reduce the blood concentration of the above-mentioned indoxyl sulfate by administering paramylon or its processed product to renal failure patients.

The specific mechanism of action is as follows.

(1) Paramylon or a processed product thereof, which is an effective main component of the inhibitor of the progression of renal failure, is porous and acts to directly adsorb indole, which is a precursor of indoxyl sulfate, in the intestine of a patient with renal failure.

In addition, paramylon or its processed product is indigestible, and in a state where indole is adsorbed, it is not absorbed into the living body, but passes through the digestive tract and is excreted in feces together with indole.

Therefore, indole is absorbed by the digestive tract, which inhibits the production of indoxyl sulfate, which is produced by the binding of sulfuric acid in the liver. Furthermore, the release of indoxyl sulfate into the blood is inhibited, so that the concentration of indoxyl sulfate in the blood can be reduced.

(2) In addition, paramylon or a processed product thereof, which is an effective main component of the inhibitor of the progression of renal failure, exerts an action that contributes to the improvement of the human intestinal environment, which is an action that the conventional spherical adsorption carbon did not have.

(2-1) In detail, when paramylon or its processed product is administered to a living body, it has the effect of activating and proliferating beneficial bacteria including bifidobacteria and lactic acid bacteria in the human intestine. Therefore, the balance-improving effect of intestinal bacteria in patients with renal failure in which harmful bacteria are usually increased and beneficial bacteria are decreased can be exhibited.

Therefore, by suppressing the increase of harmful bacteria in the intestine, the amount of indole produced by the decomposition of tryptophan contained in the protein in the food is suppressed by the harmful bacteria in the intestine, and thus the amount of production of indoxyl sulfate can be suppressed.

(2-2) In addition, when paramylon or its processed product is administered to a living body, the intestinal transit time of orally ingested food is shortened, and the amount of indole produced from tryptophan during intestinal transit is suppressed. , thus inhibiting the production of indoxyl sulfate.

According to at least the above-mentioned mechanism of action, the blood indoxyl sulfate concentration can be reduced by administering the renal failure progression inhibitor of the present embodiment to a renal failure patient.

＜＜Use＞＞

By administering the renal failure progression inhibitor of the present embodiment to an acute renal failure patient, it can be used to suppress the progression and progression of acute renal failure disease.

In particular, the concentration of indoxyl sulfate in patients with acute renal failure sometimes rises sharply. Cardiovascular disease and death have occurred due to the vascular barrier effect of indoxyl sulfate. Therefore, the effect of this renal failure progression inhibitor has been changed. be meaningful.

In addition, the present renal failure development inhibitor can be applied to patients with acute renal failure whether pre-renal, renal or post-renal, and can be used for acute renal failure in the onset, oliguria, diuretic, and recovery phases of acute renal failure. It can be applied to patients of all periods.

In addition, the present renal failure development inhibitor can also be applied to patients suffering from the above-mentioned diseases listed as causes of acute renal failure and their high-risk patients (original text: Preparatory Army), and can be used as a preventive agent for acute renal failure.

By administering the renal failure progression inhibitor of the present embodiment to a chronic renal failure patient, it can be used to inhibit the progression and progression of chronic renal failure disease.

In addition, the present renal failure progression inhibitor can be applied to patients with chronic renal failure regardless of stages 1 to 5 of CKD, and can also be applied to patients who have received kidney transplantation. For example, in patients with renal failure in the first to fourth stages of CKD in the storage period, it has the effect of delaying the time period until the introduction of dialysis.

In addition, the present renal failure progression inhibitor can also be applied to patients with diseases such as diabetic nephropathy, chronic nephritis (chronic glomerulonephritis), and nephrosclerosis, which are listed as causes of chronic renal failure, and their high-risk individuals, and Can be used as a preventive agent for chronic renal failure.

The renal failure progression inhibitor of the present embodiment can be utilized as a pharmaceutical composition containing the renal failure progression inhibitor, a composition such as a food composition, or the like.

(pharmaceutical composition)

In the field of medicine, paramylon in an amount capable of effectively exhibiting the effect of inhibiting the development of renal failure, that is, the effect of reducing the blood concentration of indoxyl sulfate, is formulated with pharmaceutically acceptable carriers and additives, thereby providing A pharmaceutical composition for this effect. The pharmaceutical composition may be a drug or a quasi-drug.

In particular, it is desirable to prepare a mixture with lactic acid bacteria, bifidobacterial lactic acid bacteria, butyric acid bacteria, etc. having an intestinal regulating effect, and the effect of the present renal failure progression inhibitor is synergistically improved. Specifically, by suppressing the proliferation of harmful intestinal bacteria such as coliforms by the above-mentioned intestinal regulating action, the effect of reducing putrefaction products produced by harmful intestinal bacteria can be exhibited synergistically.

In addition, it is desirable to form a mixture with a pH adjuster for lowering the pH (hydrogen ion concentration index) in the intestine in order to suppress the proliferation of harmful bacteria in the intestine, and the effect of the present renal failure progression inhibitor is synergistically enhanced.

The pharmaceutical composition is suitable for both internal use and external use. Therefore, the pharmaceutical composition can be used in preparation forms such as injections such as oral administration, intravenous injection, subcutaneous injection, intradermal injection, intramuscular injection and/or intraperitoneal injection, transmucosal preparations, and transdermal preparations.

The dosage form of the pharmaceutical composition can be appropriately set according to the application form, and examples include solid preparations such as tablets, granules, capsules, powders, and powders; liquid preparations such as liquid preparations and suspensions; Glue and other semi-solid agents.

(food composition)

In the food field, a food composition having this effect can be provided by incorporating an effective amount of paramylon capable of exerting an effect of inhibiting the development of renal failure in a living body as a food material in various foods.

In particular, it is desirable to prepare a food in combination with lactic acid bacteria, bifidobacterial acid bacteria, butyric acid bacteria, etc. having an intestinal regulating effect, or a food combined with a pH adjuster, and the effect of the present renal failure development inhibitor is synergistically improved.

That is, the present invention can provide a food composition exhibiting an effect of inhibiting the development of renal failure and the like in the food field. As the food composition, in addition to general foods, foods for specific health uses, nutritional functional foods, foods showing functionality, foods for hospital patients, supplements, and the like can be exemplified. In addition, it can also be used as a food additive.

Examples of the food composition include seasonings, processed meat products, processed agricultural products, beverages (soft drinks, alcoholic drinks, carbonated drinks, milk drinks, fruit juice drinks, tea, coffee, nutritional drinks, etc.), powder drinks ( Powdered juice, powdered soup, etc.), concentrated beverages, desserts (candy, cookies, biscuits, chewing gum, gummy (original: グミ), chocolate, etc.), bread, cereals, etc. In addition, in the case of a food for specific health use, a nutritional functional food, a food expressing a function, or the like, it may be in the form of a capsule, a lozenge, a syrup, a granule, a powder, or the like.

Here, a food for specific health use refers to a food containing a health functional ingredient that affects physiological functions and the like, and can mean a food suitable for a specific health use with the permission of the Director-General of the Consumer Affairs Agency. In the present invention, it refers to foods sold for specific health care purposes such as inhibiting the development of renal failure, preventing and improving renal failure, preventing and improving uremia, and inhibiting the production of indoxyl sulfate in vivo.

In addition, the nutritive functional food refers to a food that is used to supplement nutritional components (vitamins, minerals), and is a food that expresses the function of the nutritional components. Since it is sold as a nutritional functional food, it needs to be within the range of the upper limit value and the lower limit value of the amount of nutrients contained in the daily intake reference amount, which not only needs to express the nutritional function, but also needs to Notes are displayed, etc.

In addition, a functional food shows the functional food based on a scientific basis within the responsibility of a business operator. Report information on safety and functional basis to the Commissioner of the Consumer Affairs Agency prior to sale.

Among the above, the present invention contains paramylon or its processed product as an active ingredient, and can be used as a specific health food for inhibiting the progression of renal failure, kidney failure patients, and animals other than humans suffering from renal failure. Nutritional functional food for suppressing progression of failure, and functional food for suppressing progression of renal failure.

In addition, the present invention contains paramylon or a processed product thereof as an active ingredient, and can be used as an active ingredient in a living body, such as a person before renal failure, a person at high risk of renal failure, a person before diagnosis or treatment of renal failure, Specific health food for prevention of renal failure, nutritional functional food for prevention of renal failure, and functional food for prevention of renal failure for animals other than humans.

＜＜Usage and dosage＞＞

As the usage of the renal failure progression inhibitor of the present embodiment, oral administration to renal failure patients is suitable, and oral administration to chronic renal failure patients is preferable.

Here, the chronic renal failure patient may be a patient receiving dialysis therapy, a patient before receiving dialysis therapy, or a patient after receiving kidney transplantation. More preferred are chronic renal failure patients undergoing dialysis therapy. More preferably, it is a 50- to 70-year-old chronic renal failure patient receiving dialysis therapy, preferably a male patient.

The usage of the renal failure progression inhibitor of the present embodiment is suitable for oral administration of 3 to 15 g of paramylon or a paramylon-processed product per day to renal failure patients. Oral administration is preferably carried out at 3 to 9 g per day, more preferably 6 g per day. In addition, a predetermined amount of paramylon or paramylon-processed product is preferably administered once a day, preferably several times a day, and more preferably three times a day.

In addition, the use of the renal failure development inhibitor is suitable for continuous administration to renal failure patients, preferably continuous administration for 9 weeks or more.

In addition, as the dosage form of the renal failure progression inhibitor, solid preparations such as tablets, granules, capsules, powders, and powders are preferred, and capsules or powders are more preferred.

The usage of the renal failure progression inhibitor of the present embodiment is preferably administered to a renal failure patient before or after a meal.

In addition, the use of the renal failure development inhibitor is suitable to be administered alone to renal failure patients without simultaneous administration with other agents or the like. The administration is preferably carried out with a predetermined period of time before and after the administration of other drugs.

As a comparison object, in the case of spherical adsorbent carbon, which is a conventional inhibitor of the development of renal failure, it is usually necessary to take 6g (30 capsules for a 200mg capsule, or 2g/pack of granules for a 200mg capsule) three times a day. for 3 packs). This dosage is much larger than the dosage of ordinary medicines, and even if it is a granule preparation, it is difficult to take it due to a feeling of brushing (original: ジャリジャリ) in the oral cavity. In addition, there are side effects such as abdominal distention and constipation, so the burden on the patient is considerable.

On the other hand, in the case of the paramylon or paramylon processed product of the present invention, even if it is the same as 6 g per day, 24 capsules or powders as 250 mg capsules can be divided into 3 doses to be taken, and the number of capsules can be reduced. number of doses. For example, when it is a powder, the texture is also good, and there are no side effects such as abdominal distention and constipation. Therefore, the pain and burden of the renal failure patient are reduced, and the patient can easily take it for a long time.

＜Therapeutic agent for renal failure and preventive agent for renal failure＞

In this embodiment, the inhibitory agent for the development of renal failure has been mainly described, but it can also be used as an active ingredient containing paramylon or a processed product thereof, and a patient with renal failure or an animal other than a human suffering from renal failure as an active ingredient. A therapeutic agent for renal failure in a subject.

The renal failure therapeutic agent reduces the blood indoxyl sulfate concentration in the living body to the normal range by administering paramylon to the living body.

Specifically, the synergistic enhancement of the present renal failure progression inhibitor can be provided by blending paramylon in an amount that can effectively reduce the concentration of indoxyl sulfate in blood together with pharmaceutically acceptable carriers and additives. Effective renal failure therapeutic agent.

In addition, it can also be used as an active ingredient containing paramylon or a processed product thereof and used in living organisms, such as people before renal failure, people at high risk of renal failure, people before diagnosis or treatment of renal failure, A renal failure preventive agent for other animals.

The renal failure preventive agent controls the blood indoxyl sulfate concentration in the living body within the normal range by administering paramylon to the living body.

<Therapeutic and preventive agents other than indoxyl sulfate production inhibitors>

The present invention can also be used as an indoxyl sulfate production inhibitor that contains paramylon or a processed product thereof as an active ingredient and reduces the blood concentration of indoxyl sulfate produced in vivo.

Indoxyl sulfate production inhibitor functions not only as an inhibitor of the progression of renal failure, a therapeutic agent, or a preventive agent, but also as a disease related to renal failure, ie, nephritis (filamentous nephritis), nephropathy (diabetic nephropathy, IgA nephropathy) ), a therapeutic or preventive agent for cardiovascular disease, heart failure, myocardial infarction, stroke, etc. In addition, it functions as a preventive or therapeutic agent for diseases that are improved by reducing the blood indoxyl sulfate concentration.

In addition, the present invention can also be used as a uremic therapeutic agent administered to a uremic patient, and a therapeutic agent for a cardiovascular system disease associated with chronic kidney disease (CKD) administered to a chronic kidney disease (CKD) patient.

In addition, it can also be used as a preventive or therapeutic agent for diseases related to renal failure such as nephritis (filamentous nephritis), nephropathy (diabetic nephropathy, IgA nephropathy), cardiovascular disease, heart failure, myocardial infarction, stroke, etc., Auxiliary health status micromodulator after treatment.

Example

<Example>

Paramylon derived from Euglena was prepared (manufactured) by the following procedure.

Euglena fines powder (manufactured by Euglena Co., Ltd.) was added to distilled water, and the mixture was stirred at room temperature for 2 days. It was subjected to ultrasonic treatment to destroy the cell membrane, and the crude paramylon particles were recovered by centrifugation. The recovered paramylon particles were dispersed in a 1% sodium dodecyl sulfate aqueous solution, treated at 95°C for 2 hours, and centrifuged again, and the recovered paramylon particles were dispersed into 0.1% dodecane in aqueous sodium sulfate solution at 50°C for 30 minutes. By this operation, lipids and proteins were removed, followed by washing with acetone and ether, and drying at 50°C to obtain purified paramylon particles.

The prepared paramylon was contained in a known capsule in the form of a solid preparation to obtain a renal failure progression inhibitor.

<Test Example 1 Test of administration of a renal failure development inhibitor to patients with chronic renal failure>

Using the renal failure development inhibitor of the Example, a human clinical test of the renal failure development inhibitory effect was carried out.

The subjects of this trial were 48 chronic renal failure patients (16 males and 32 females) aged 40-88 who received dialysis therapy, with an average age of 67.81 years and an average dialysis treatment experience of 9.88 years.

The subjects were randomly divided into a paramylon group (29 subjects) to which the renal failure development inhibitor was administered and a control group (19 subjects) to which the renal failure development inhibitor was not administered. The subjects in the paramylon group were allowed to orally ingest the renal failure development inhibitor of the example alone between meals at 2 g (8 capsules or powders) three times a day each time. In particular, oral ingestion is performed with a predetermined period of time before and after administration of other drugs. Oral ingestion continued for 9 weeks.

It should be noted that, of the 29 subjects in the paramylon group, 3 were withdrawn due to the present renal failure development inhibitor (the amount of capsules was too large to be taken orally, etc.), and 4 were withdrawn due to various circumstances. In addition, among the 19 people in the control group, 1 person was removed due to various circumstances.

Blood was collected from each test subject immediately before the start of ingestion of the renal failure development inhibitor and 9 weeks after the start of ingestion. Using the collected whole blood, the concentration of indoxyl sulfate in the blood was measured, and the amount of change was monitored. A specific measurement method is as follows.

(1) Sample pretreatment

Trichloroacetic acid (4% TCA) was added to 10 μL of the test subject's serum, and after stirring, centrifugation was performed, and the supernatant was collected into a sample bottle for analysis to prepare a serum sample (deproteinized). Serum samples were diluted 10-fold by this treatment.

The standard indoxyl sulfate storage solution was added to 10 μL of serum from healthy people so that the final concentrations of 0.0 (no addition) and 5.0 (μg/ml) were added, and the above-mentioned protein removal treatment was performed to prepare an external standard calibration curve (Original: External standard gauge) with samples 1, 2.

(2) HPLC quantitative analysis based on external standard calibration curve method

Using a high-performance liquid chromatography (HPLC) apparatus (L-2000 manufactured by Hitachi High-Technologies), 2.5 μL of each of the supernatants of the above-mentioned standards 1 and 2 were separated and analyzed, and the concentration (μg/ml) was calculated by the external standard calibration curve method. .

(3) Sample analysis

The above serum samples were analyzed in the stated order.

External standard method calibration curve The first and last two calibration curves were prepared by fitting the analysis results of samples 1 and 2 by the least squares method, and the concentration (µg/ml) of indoxyl sulfate in the serum sample was determined.

The final blood indoxyl sulfate concentration was obtained by multiplying the analysis result of the serum sample by the dilution factor of 10. In addition, the serum sample of a healthy person was prepared as a blank, and the analysis result of the serum sample of a test subject was added up in advance.

The age (years), dialysis treatment experience (years), sex, measurement results of blood indoxyl sulfate concentration (μg/ml) before and after the start of ingestion of each test subject were divided into a control group and a paramylon group, and the The two groups were compared separately.

Figure 1 shows a graph comparing the average change (average decrease) in blood indoxyl sulfate concentration before and after the start of ingestion of the control group (22 persons) and the paramylon group (18 persons) by analyzing the above measurement results. Show.

In addition, the subjects were limited to a group (18 people) aged 50 years or older and 70 years or younger, and the blood indoxyl sulfate before and after the intake of the control group (8 people) and the paramylon group (10 people) was started. Fig. 2 shows a graph comparing the average amount of change in concentration (average amount of decrease).

In addition, the subjects were limited to the male group (22 subjects), and the average change amount (mean decrease in blood indoxyl sulfate concentration) before and after the start of ingestion of the control group (6 subjects) and the paramylon group (16 subjects) was determined. A comparison chart is shown in Figure 3.

According to FIG. 1 , the blood indoxyl sulfate concentration of all the patients in the paramylon group was significantly decreased during the 9-week administration period (P<0.05 by t-test). On the other hand, there was no significant change in the blood indoxyl sulfate concentration of all the patients in the control group.

In addition, according to FIG. 2 , in the group aged 50 to 70 years old, the blood indoxyl sulfate concentration of the patients in the paramylon group was significantly decreased (t-test, P<0.05). On the other hand, there was no significant change in the blood indoxyl sulfate concentration of the patients in the control group.

In addition, according to FIG. 3 , in the male population, the blood indoxyl sulfate concentration of the patients in the paramylon group was significantly decreased (by t-test, P<0.05). On the other hand, there was no significant change in the blood indoxyl sulfate concentration of the patients in the control group.

(Investigation of Test Example 1)

According to the results of Test Example 1, the concentration of indoxyl sulfate in blood was significantly decreased by continuous administration of the renal failure progression inhibitor of Example to 40 to 88-year-old chronic renal failure patients undergoing dialysis therapy for 9 weeks. .

In particular, the renal failure progression inhibitor was orally administered to the patient 3 times a day and 2 g (8 capsules or powders) each time between meals alone, and the administration was performed with a prescribed time before and after the administration of other drugs. , thereby significantly reducing the concentration of indoxyl sulfate in the blood.

In addition, in patients with chronic renal failure aged 40 to 88 who received dialysis therapy, the concentration of indoxyl sulfate in blood was significantly reduced by administering a renal failure progression inhibitor to patients aged 50 to 70 years in particular. .

In addition, in patients with chronic renal failure aged 40 to 88 years who received dialysis therapy, the concentration of indoxyl sulfate in blood was significantly decreased by administering a renal failure progression inhibitor to male patients in particular.

In addition, in general, when conventional spherical adsorbent carbon is continuously administered to renal failure patients, it is difficult for about half of the patients to take oral administration, and most of them are withdrawn due to reasons such as abdominal distention, constipation, etc. side effects. In the case of the development inhibitor, among the 29 patients in the paramylon-administered group, only 3 patients were released on the grounds of this renal failure development inhibitor.

According to the actual doctor's interview, as the subject's impression, it was found that it was easier to take than the conventional spherical adsorption charcoal, and the symptoms such as abdominal distention and constipation were not improved.

Furthermore, the following doctor's diagnosis results were obtained: This Test Example 1 was carried out on the premise that the subject was not informed of the effect of the present inhibitor of the development of renal failure, so if the subject was informed of the effect of the present inhibitor of the development of renal failure in advance , there should be fewer patients disengaged.

In particular, the following doctor's diagnosis results were obtained: Generally, renal failure patients undergoing dialysis treatment are prone to constipation under the restriction of water intake, and if the patient (subject) is informed in advance of one of the effects of this renal failure development inhibitor That is, the improvement effect of the intestinal environment, the effect of improving constipation, which is an effect not possessed by the conventional spherical adsorption carbon, can be confirmed, and therefore, there will be fewer patients who are detached.

Based on the above, it can be seen that the renal failure progression inhibitor of the Example suppresses the progression of renal failure disease in chronic renal failure patients receiving dialysis therapy by reducing the blood indoxyl sulfate concentration in vivo.

Furthermore, compared with the conventional renal failure development inhibitor, namely spherical adsorbent carbon, it is easier to take and less likely to cause side effects such as abdominal distention and constipation, so that chronic renal failure patients are easy to take orally for a long time.

<Test example 2 Indole adsorption capacity test>

A test for confirming the indole adsorption capacity of the paramylon prepared in the Example was conducted.

A specific method is as follows.

(1) Preparation of calibration curve

Indole solutions with concentrations of 0, 10, 20, 30, 40, and 50 ng/ml were prepared, respectively, and the absorbance at the excitation wavelength of 342 nm was measured using a fluorophotometer (F-2500, manufactured by Hitachi High-Technologies). Calibration curve for the relationship of absorbance.

(2) Determination of indole residual concentration after paramylon mixing

Paramylon was added to the indole solution at a concentration of 30 ng/ml in an amount to achieve 2% volume, and mixed for 1 minute using a vortex mixer. Then, it was centrifuged for 1 minute at 6200 rpm using a centrifuge to precipitate paramylon, and the supernatant was collected as a sample.

As specific samples, sample 1: supernatant liquid collected by centrifugation immediately after the above mixing, sample 2: supernatant liquid collected after mixing and allowed to stand for 10 minutes, and then centrifuged and collected, sample 3: mixed and allowed to stand still The supernatant liquid collected after being centrifuged for 30 minutes, Sample 4: The supernatant liquid collected after being mixed and allowed to stand for 60 minutes, then centrifuged and collected, Sample 5: After being mixed and allowed to stand for 1440 minutes (24 hours) A total of 5 samples of the supernatant liquid recovered by centrifugation were measured for absorbance at an excitation wavelength of 342 nm using a fluorophotometer.

On the other hand, paramylon was added to purified water in an amount of 2% of the volume, mixed for 1 minute, centrifuged to precipitate paramylon, the supernatant was collected as a blank, and the excitation wavelength was measured. Absorbance at 342 nm.

From the absorbance values measured in Samples 1 to 5, the absorbance values measured by the blank solution were subtracted, and the residual indole concentrations in the indole solutions were calculated respectively, thereby confirming the time-lapse of the indole adsorption capacity by paramylon. sexual changes.

As a test result, a graph showing the relationship between the standing time after paramylon mixing and the residual indole concentration is shown in FIG. 4 .

As can be seen from FIG. 4 , when paramylon was mixed with the indole solution, the residual concentration of indole in the indole solution decreased.

In addition, it was found that after mixing paramylon in the indole solution, the longer the standing time was, the lower the indole residual concentration in the indole solution was.

(Investigation of Test Example 2)

From the results of Test Example 2, it was found that when paramylon was mixed with the indole solution, the indole contained in the indole solution was adsorbed by the paramylon.

In addition, it can be seen that after mixing paramylon in the indole solution, the longer the standing time, the greater the indole adsorption capacity of paramylon.

From the above, it was found that the paramylon of the Examples exhibited an action of adsorbing indole in the intestine of a human, especially a patient with renal failure.

Furthermore, it was found that the concentration of indoxyl sulfate in the blood can be reduced by reducing the absolute amount of indole, which is a precursor of indoxyl sulfate, and inhibiting the release of indoxyl sulfate into the blood.

<Test Example 3 Proliferation Confirmation Test of Intestinal Beneficial Bacteria>

The paramylon prepared in the Example was orally ingested to rats, and a test for confirming the improvement effect of paramylon on the intestinal environment, specifically, a test for measuring the occupancy rate of beneficial bacteria in the intestinal flora was conducted.

For the test, 15 3-week-old Wistar male rats (CLEA Co., Ltd., Japan) were used. Feed and drinking water (distilled water) were freely ingested.

After 1 week of preparatory feeding, the rats were divided into 3 groups of 5 rats and fed for 4 weeks. The feed was prepared based on purified feed (AIN-93N; Japan CLEA Co., Ltd.), and the control group was orally ingested with the feed from which cellulose was removed. In addition, the cellulose group ingested a feed supplemented with 5% cellulose, and the paramylon group ingested a feed supplemented with 5% paramylon prepared in Example instead of cellulose (refer to the feed composition shown in Table 1 below) .

Table 1

※The total value is rounded up to 100.

In the test, Lactobacillus and Bifidobacterium, which are classified as beneficial bacteria, Clostridium, which is classified as harmful bacteria, and Prevotella, which is classified as opportunistic pathogenic bacteria, in the intestinal flora of each rat. The respective occupancy rates of the genera Prevotella and Bacteroides were measured for each bacterial group. In addition, the ratio of beneficial bacteria and harmful bacteria was obtained.

Specifically, with respect to the intestinal flora in the cecum of each rat, the base arrangement of the 16S rRNA portion derived from the flora was analyzed based on a known method.

(Nagashima K, et al. (2003) "Application of New Primer-Enzyme Combinations to Terminal Restriction Fragment Length Polymorphism Profiling of Bacterial Populations in Human Feces" Appl Environ Microbiol, 69, 1251-1262.)

As a test result, a graph comparing the occupancy rates of intestinal flora in the cecum between the control group, the cellulose group, and the paramylon group is shown in FIG. 5 . In addition, a graph comparing the ratio of beneficial bacteria/harmful bacteria in the cecum is shown in FIG. 6 .

According to FIG. 5 , the occupancy rate of beneficial bacteria was higher in the paramylon group and the cellulose group than in the control group. In terms of the occupancy rate of harmful bacteria, no significant difference was observed between the three groups. In terms of the occupancy rate of opportunistic pathogens, the paramylon group was higher than the other groups.

In addition, according to FIG. 6 , the paramylon group was higher than the other groups in terms of the ratio of beneficial bacteria to harmful bacteria.

(Investigation of Test Example 3)

According to the results of Test Example 3, in the paramylon group and the cellulose group rats that continuously ingested paramylon and cellulose, which are insoluble dietary fibers, the occupancy rate of beneficial bacteria in the intestinal flora was high. In addition, in the paramylon group rats, the ratio of beneficial bacteria to harmful bacteria in the intestinal flora showed higher values than the other groups.

That is, paramylon has the effect of improving the intestinal environment, and has the effect of improving the intestinal environment equal to or higher than that of cellulose.

From the above, it was found that when paramylon is orally administered to a living body, it has the effect of activating and proliferating beneficial bacteria including bifidobacteria and lactic acid bacteria in the intestine, and also has the effect of improving the balance of intestinal bacteria.

As a result, it was found that by suppressing the increase of harmful bacteria in the intestine, the amount of indole produced by the decomposition of tryptophan contained in the protein in the food by the harmful bacteria in the intestine is suppressed, and the amount of production of indoxyl sulfate is suppressed. .

In particular, in the case of the conventional spherical adsorbent carbon, it was found that the administration could not be performed because of the risk of excretion disturbance in renal failure patients with impaired digestive tract passage. The patient can also administer the drug without obstructing excretion.

<Test Example 4 Intestinal transit time measurement test of orally ingested food>

The paramylon prepared in the Example was orally ingested to rats, and a test for confirming the improvement effect of paramylon on the intestinal environment, specifically, a test for measuring the intestinal transit time of the orally ingested paramylon-containing feed was performed.

In the test, as in Test Example 2, 15 3-week-old Wistar male rats were used. Feed and drinking water (distilled water) were freely ingested.

After 1 week of preparatory feeding, the rats were divided into 3 groups of 5 rats and fed for 4 weeks. The control group was ingested orally with cellulose-removed feed. In addition, the cellulose group ingested a feed supplemented with 5% cellulose, and the paramylon group ingested a feed supplemented with 5% paramylon prepared in Example instead of cellulose (refer to the feed composition shown in Table 1 above).

In the test, rats were ingested at 18:00 on the second week (day 13) and the fourth week (day 27) from the start of feeding, and 5% magenta pigment was added to the diet, and the amount of feces until red feces was observed was measured. time. Confirmation is performed every 1 hour from 0 hours to 3 hours, and every 30 minutes after 3:00.

During the rearing period, the animals were fasted from 9:00 to 18:00 according to the intestinal transit time measured in the second and fourth weeks, and they were allowed to ingest freely at other times.

As a test result, the graph which compares the intestinal transit time of the control group, the cellulose group, and the paramylon group at the 2nd week and the 4th week from the feeding start, respectively, is shown in FIG. 7 .

According to FIG. 7 , no significant difference was observed between the control group, the cellulose group, and the paramylon group in the intestinal transit time measured in the second week.

The paramylon group was the earliest in terms of intestinal transit time measured at week 4. The paramylon group and the cellulose group were significantly earlier than the control group. No significant difference was recognized between the paramylon group and the cellulose group.

(Investigation of Test Example 4)

According to the results of Test Example 4, by continuously administering paramylon to the rat for 4 weeks, it was observed that the rat's intestinal transit time was shorter than that of the control group.

In addition, by the continuous administration of paramylon, a reduction in the intestinal transit time equivalent to that of the continuous administration of cellulose was observed. That is, it was found that paramylon has an effect of improving the intestinal environment equivalent to that of cellulose.

From the above, when paramylon is orally administered to a living body, the intestinal transit time of the orally ingested food is shortened.

As a result, it was found that the production amount of indole produced from tryptophan during intestinal passage was suppressed, and the production amount of indoxyl sulfate was also suppressed.

<Test Example 5 Weight Measurement Test of Feces>

The paramylon prepared in the Example was orally ingested to rats, and a test for confirming the improvement effect of paramylon on the intestinal environment, specifically, a test for measuring the weight of feces excreted after oral ingestion of paramylon was performed.

In the test, 15 3-week-old Wistar male rats were used in the same manner as in Test Examples 2 and 3. Feed and drinking water (distilled water) were freely ingested.

After 1 week of preparatory feeding, each group of 5 rats was divided into 3 groups: control group, cellulose group and paramylon group, and were fed for 4 weeks.

In the test, feces were collected on 3 days (11th to 13th days) in the second week of rearing and 3 days before slaughter (25th to 27th days) in the fourth week of rearing, and the dry weight and moisture content of the feces were measured. . In addition, the moisture content of stool was weighed after drying stool at 100 degreeC for 24 hours.

As test results, graphs comparing the weight and water content of feces in the second and fourth weeks after the feeding of the control group, the cellulose group, and the paramylon group, respectively, are shown in FIGS. 8 and 9 .

According to FIG. 8 , the weights of stools measured in the second and fourth weeks were significantly larger in the paramylon group and the cellulose group than in the control group (by t-test, P<0.05). No significant difference was recognized between the paramylon group and the cellulose group.

According to FIG. 9 , the paramylon group was significantly larger than the control group and higher than the cellulose group in terms of the water content of the feces measured at the 4th week. (P<0.05 using t-test).

(Investigation of Test Example 5)

From the results of Test Example 5, when paramylon was continuously administered to the rat for 4 weeks, it was observed that the weight and water content of the feces excreted by the rat were increased compared to the control group.

In addition, by continuous administration of paramylon, an increase in fecal volume equivalent to that of continuous administration of cellulose was observed. That is, it was found that paramylon has an effect of improving the intestinal environment equivalent to that of cellulose.

From the above, it was found that when paramylon was administered to a living body, the weight and water content of stool after oral ingestion increased.

In general, it has been reported that if the weight of feces increases and the water retention of feces increases, the intestinal transit time of food is shortened (and Chuan Guizi, the fecal volume and digestive tract transit time of so-called dietary fiber beverages in rats and human The influence of the convenience caused by the communication, Iwate University Faculty of Education Research Annual Report, 55, 111-118 (1995)).

Therefore, it was found that the production amount of indole produced from tryptophan during intestinal passage was suppressed, and the production amount of indoxyl sulfate was suppressed.

Claims (4)

1. Application of paramylon in preparation of inhibitor for renal failure development is provided.

2. Application of paramylon in preparation of prophylactic agent for renal failure is provided.

3. Application of Euglena starch in preparation of therapeutic agent for uremia is provided.

4. The use according to any one of claims 1 to 3, wherein the inhibitor, prophylactic or therapeutic agent is a substance for inhibiting the production of indoxyl sulfate.

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